U.S. patent application number 15/653814 was filed with the patent office on 2018-01-25 for bezafibrate for the treatment of cancer.
The applicant listed for this patent is Cipla Limited. Invention is credited to Jeevan Ghosalkar, Kalpana Joshi, Geena Malhotra.
Application Number | 20180021276 15/653814 |
Document ID | / |
Family ID | 60989426 |
Filed Date | 2018-01-25 |
United States Patent
Application |
20180021276 |
Kind Code |
A1 |
Malhotra; Geena ; et
al. |
January 25, 2018 |
BEZAFIBRATE FOR THE TREATMENT OF CANCER
Abstract
Methods of treating renal cancer, including renal cell
carcinoma, using bezafibrate are disclosed herein. Bezafibrate can
be administered as a monotherapy or as part of a comprehensive
treatment program, which can also include administration with other
anti-cancer drugs, surgical treatments or exposure to ionizing
radiation.
Inventors: |
Malhotra; Geena; (Mumbai,
IN) ; Joshi; Kalpana; (Maharashtra, IN) ;
Ghosalkar; Jeevan; (Thane, IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Cipla Limited |
Mumbai |
|
IN |
|
|
Family ID: |
60989426 |
Appl. No.: |
15/653814 |
Filed: |
July 19, 2017 |
Current U.S.
Class: |
424/133.1 |
Current CPC
Class: |
A61K 45/06 20130101;
A61K 9/0053 20130101; A61K 31/195 20130101; A61K 31/404 20130101;
A61K 2300/00 20130101; A61K 2300/00 20130101; A61K 31/192 20130101;
A61K 9/2054 20130101; A61K 9/4858 20130101; A61K 31/404 20130101;
A61K 31/195 20130101 |
International
Class: |
A61K 31/192 20060101
A61K031/192; A61K 31/404 20060101 A61K031/404; A61K 9/00 20060101
A61K009/00; A61K 45/06 20060101 A61K045/06 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 20, 2016 |
IN |
201621024898 |
Claims
1. A method for the treatment of renal cell carcinoma in a patient,
the method comprising administering to said patient bezafibrate, or
a pharmaceutically acceptable salt or prodrug thereof, in an amount
effective to treat renal cell carcinoma.
2. The method according to claim 1, wherein the renal cell
carcinoma comprises treat conventional (clear-cell) renal cell
carcinoma, papillary renal cell-carcinoma, chromophobe renal
carcinoma, onco-cytoma, or collecting-duct carcinoma.
3. The method according to claim 1, wherein the renal cell
carcinoma comprises Stage I, Stage II, Stage III or Stage IV renal
cell carcinoma.
4. The method according to claim 1, wherein bezafibrate is
administered in combination with at least one other cancer
therapy.
5. The method according to claim 4, wherein the additional cancer
therapy comprises surgery, chemotherapy, immunotherapy, or ionizing
radiation.
6. The method according to claim 5, wherein the additional cancer
therapy comprises administering at least one additional agent
comprising a nucleoside analogue, antifolate, antimetabolite,
topoisomerase I inhibitor, anthracycline, podophyllotoxin, taxanes,
vinca alkaloid, alkylating agent, platinum compound, proteasome
inhibitor, nitrogen mustards, oestrogen analogue, monoclonal
antibody, tyrosine kinase inhibitor, mTOR inhibitor, retinoid,
immunomodulatory agent, histone deacetylase inhibitor, or other
kinase inhibitor.
7. The method according to claim 6, wherein the at least one
additional agent comprises bevacizumab, axitinib, sorafenib,
sunitinib, everolimus, temsirolimus, pazopanib, or lenvatinib.
8. The method according to claim 6, wherein the at least one
additional agent comprises sunitinib.
9. The method according to claim 6, wherein the at least one
additional agent is administered simultaneously with
bezafibrate.
10. The method according to claim 6, wherein the at least one
additional agent is administered sequentially with bezafibrate.
11. The method according to claim 6, wherein the at least one
additional agent is administered separately from bezafibrate.
12. The method according to claim 4, wherein the bezafibrate is
administered prior to treatment with ionizing radiation
13. The method according to claim 4, wherein the bezafibrate is
administered subsequent to treatment with ionizing radiation
14. The method according to claim 4, wherein the bezafibrate is
administered prior to surgical treatment for renal cell
carcinoma.
15. The method according to claim 4, wherein the bezafibrate is
administered subsequent to surgical treatment for renal cell
carcinoma.
16. A pharmaceutical composition comprising bezafibrate and at
least one additional renal cell carcinoma therapeutic agent.
17. A kit comprising bezafibrate and at least one additional renal
cell carcinoma therapeutic agent.
18. The kit according to claim 17, wherein the bezafibrate is
provided in a pharmaceutical composition suitable for oral
administration.
19. The kit according to claim 17, wherein the bezafibrate is
provided in a pharmaceutical composition suitable for parenteral
administration.
20. The kit according to claim 17, wherein the bezafibrate and at
least one additional therapeutic agent are provided in separate
pharmaceutical compositions.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application claims the benefit of Indian Application
201621024898, filed on Jul. 20, 2016, the contents of which are
incorporated herein in its entirety.
FIELD OF THE INVENTION
[0002] The invention is directed to the treatment of cancer,
especially renal cell carcinoma, using bezafibrate, optionally in
combination with one or more additional cancer therapeutics.
BACKGROUND
[0003] Renal cell carcinoma (RCC, also known as hypernephroma) is a
kidney cancer that originates in the lining of the proximal
convoluted tubule, the very small tubes in the kidney that
transport GF (glomerular filtrate) from the glomerulus to the
descending limb of the nephron. RCC is the most common type of
kidney cancer in adults, responsible for approximately 80% of
cases. It is also known to be the most lethal of all the
genitourinary tumors. Initial treatment is most commonly a radical
or partial nephrectomy and remains the mainstay of curative
treatment. Where the tumor is confined to the renal parenchyma, the
five year survival rate is 60-70%, but this is lowered considerably
once metastases have spread. It is relatively resistant to
radiation therapy and chemotherapy, although some cases respond to
immunotherapy.
[0004] Renal-cell carcinoma affects approximately 150,000 people
worldwide each year, causing close to 78,000 deaths annually, and
its incidence seems to be increasing. RCC is not a single entity,
but rather comprises the class of tumors of renal epithelial
origin. Extensive histological and molecular evaluation has
resulted in the development of a consensus classification of
different RCC subtypes: (i) conventional (clear-cell) renal cell
carcinoma; (ii) papillary renal cell-carcinoma; (iii) chromophobe
renal carcinoma; (iv) onco-cytoma; (v) collecting-duct carcinoma.
Although most cases of RCC seem to occur sporadically, an inherited
predisposition to renal cancer accounts for 1-4% of cases and could
involve the same genes that cause sporadic renal cancer. Over the
past two decades, studies of families with inherited RCC have laid
the groundwork for the identification of seven hereditary renal
cancer syndromes, and the predisposing genes for five of these have
been identified. The surprisingly diverse nature of these genes
implicates various mechanisms and biological pathways in RCC
tumorigenesis.
[0005] RCC has been conventionally treated using surgery, radiation
therapy, immunotherapy, and molecular-targeted therapy. Surgical
resection remains the only known effective treatment for localized
renal cell carcinoma, and it also is used for palliation in
metastatic disease. Targeted therapy and immunomodulatory agents
are considered standard of care in patients with metastatic
disease.
[0006] Options for chemotherapy and endocrine-based approaches are
limited, and no hormonal or chemotherapeutic regimen is accepted as
a standard of care. Objective response rates with chemotherapy,
either single-agent or combination, are usually lower than 15%.
Therefore, various therapies have been evaluated.
[0007] The first agent, approved in late 2005, was sorafenib, after
showing improvement in the second-line setting for progression-free
survival (PFS) versus placebo. Shortly thereafter, sunitinib was
approved following a large phase III trial that also demonstrated
improvement in PFS versus interferon-.alpha. (INF.alpha.) in the
first-line setting. The next agent approved was the mechanistic
target of rapamycin (serine/threonine kinase) (mTOR) inhibitor,
temsirolimus, which was evaluated as a first-line therapy against
INF.alpha. in patients, most of whom had poor-risk disease. This
trial demonstrated an improvement in overall survival (OS) in
patients receiving temsirolimus. Combination of temsirolimus and
INF.alpha. showed no advantages over the mTOR inhibitor alone.
Meanwhile, everolimus was the second mTOR inhibitor approved after
second-line therapy showed improvement in PFS versus placebo in a
clinical trial. Pazopanib and axitinib are the two newer tyrosine
kinase inhibitors and were recently approved for treatment of
metastatic RCC. Patients taking pazopanib exhibited improved PFS
versus those taking placebo both in the first-line setting and for
cytokine-refractory disease. Axitinib was studied against sorafenib
as a second-line agent and demonstrated improved PFS, while patient
preference studies with pazopanib suggested improved tolerability.
Yet another class of drug, an anti-PD-1 checkpoint inhibitor named
as nivolumab, has been approved for intravenous administration that
unleashes the body's immune system so that it can reject the kidney
cancer, however, the drug may cause the body to develop an immune
reaction against its own tissues thereby leading to wide range of
side effects that can be severe or life-threatening. With multiple
approved agents available, further research is yet to define the
ideal timing, sequencing, and patient profile for a given
particular agent.
[0008] Although, studies have demonstrated the general tolerability
of targeted agents, at most occasions, most patients with RCC
inevitably develop resistance to targeted agents after a median of
5-11 months of treatment. Combinations of targeted agents are being
evaluated, but toxicity is problematic. Several strategies have
been tested to manage the drug resistance including: Adjusting the
dose of the drug, combination therapy or switching to an
alternative agent. Moreover alternative pathways are currently
under investigation particularly targeting of RAF (Rapidly
Accelerated Fibrosarcoma), MEK (Mitogen-activated
protein/extracellular signal-regulated kinase), and the PI3K
(Phosphatidylinositol 3-kinase)/AKT (a serine/threonine kinase also
known as protein kinase B [PKB]) pathway.
[0009] Based on the information available, even though there have
been some advancements in the treatment of renal cell carcinomas,
the associated complications like the disease stage, the response
rate and the accompanying side effects potentially reduce the
patient compliance and poses issues which severely affect the
progression-free survival (PFS) and/or the overall survival (OS)
which is the ultimate treatment goal for a given therapy.
[0010] There remains a need for improved and additional methods of
treating renal cell carcinoma. There remains a need for additional
small-molecule therapeutics for the treatment of renal cancer.
SUMMARY
[0011] According to one aspect of the present invention, there is
provided a method of treating renal cell carcinoma (RCC) comprising
administration of fibrate drugs such as bezafibrate.
[0012] According to another aspect of the present invention, there
is provided a pharmaceutical composition comprising bezafibrate
with one or more pharmaceutically acceptable excipients for the
treatment of renal cell carcinoma (RCC).
[0013] According to another aspect of the present invention, there
is provided a method of treating renal cell carcinoma (RCC) by
administration of bezafibrate in combination with one or more
additional cancer treatment regimens. The cancer treatment regimen
can include administration of one or more additional therapeutic
agents, exposure to ionizing radiation, and/or surgical
interventions.
[0014] According to another aspect of the present invention, there
is provided a use of bezafibrate in combination with one or more
therapeutic agents either simultaneously, sequentially, or
separately for the treatment of renal cell carcinoma (RCC). In some
instances, the therapeutic agent can include one or more
chemotherapeutic drugs.
[0015] According to another aspect of the present invention, there
are provided pharmaceutical compositions and kits including
bezafibrate and at least one other therapeutic agent.
[0016] The details of one or more embodiments are set forth in the
descriptions below. Other features, objects, and advantages will be
apparent from the description and from the claims.
BRIEF DESCRIPTION OF THE FIGURES
[0017] FIG. 1 depicts a concentration-effect curve exhibiting
in-vitro efficacy of bezafibrate in human tumor cells in 2D
model.
[0018] FIG. 2 depicts a concentration-effect curve in-vitro
exhibiting efficacy of bezafibrate in human tumor 3D model.
[0019] FIG. 3 includes a depiction of the anti-tumor efficacy of
Bezafibrate in combination with Sunitinib against cell line
RXF1183L. Bliss index indicates the difference of Bliss neutral and
modeled T/C for each pair of conditions. Where positive values
(Bliss Index .gtoreq.0.15) indicate synergism, negative values
(Bliss Index .ltoreq.-0.15) indicate antagonism, and zero is the
neutral value.
[0020] FIG. 4 includes a depiction of the anti-tumor efficacy of
Bezafibrate in combination with Sunitinib against cell line
RXFSN12C. Bliss index indicates the difference of Bliss neutral and
modeled T/C for each pair of conditions. Where positive values
(Bliss Index .gtoreq.0.15) indicate synergism, negative values
(Bliss Index <-0.15) indicate antagonism, and zero is the
neutral value.
[0021] FIG. 5 includes a graphical representation of mean tumor
volume across the animal group populations G1 (vehicle), G2
(sunitinib), G3 (bezafibrate), and G4 (bezafibrate and
sunitinib).
[0022] FIG. 6 includes a graphical representation of mean tumor
growth inhibition (in percentage value) across the animal group
populations G1 (vehicle), G2 (sunitinib), G3 (bezafibrate), and G4
(bezafibrate and sunitinib).
[0023] FIG. 7 includes a graphical representation depicting change
in body weights across the animal group population G1 (vehicle), G2
(sunitinib), G3 (bezafibrate), and G4 (bezafibrate and
sunitinib).
DETAILED DESCRIPTION
[0024] Before the present methods and systems are disclosed and
described, it is to be understood that the methods and systems are
not limited to specific synthetic methods, specific components, or
to particular compositions. It is also to be understood that the
terminology used herein is for the purpose of describing particular
embodiments only and is not intended to be limiting.
[0025] As used in the specification and the appended claims, the
singular forms "a," "an" and "the" include plural referents unless
the context clearly dictates otherwise. Ranges may be expressed
herein as from "about" one particular value, and/or to "about"
another particular value. When such a range is expressed, another
embodiment includes from the one particular value and/or to the
other particular value. Similarly, when values are expressed as
approximations, by use of the antecedent "about," it will be
understood that the particular value forms another embodiment. It
will be further understood that the endpoints of each of the ranges
are significant both in relation to the other endpoint, and
independently of the other endpoint.
[0026] "Optional" or "optionally" means that the subsequently
described event or circumstance may or may not occur, and that the
description includes instances where said event or circumstance
occurs and instances where it does not.
[0027] Throughout the description and claims of this specification,
the word "comprise" and variations of the word, such as
"comprising" and "comprises," means "including but not limited to,"
and is not intended to exclude, for example, other additives,
components, integers or steps. "Exemplary" means "an example of"
and is not intended to convey an indication of a preferred or ideal
embodiment. "Such as" is not used in a restrictive sense, but for
explanatory purposes.
[0028] Disclosed are components that can be used to perform the
disclosed methods and systems. These and other components are
disclosed herein, and it is understood that when combinations,
subsets, interactions, groups, etc. of these components are
disclosed that while specific reference of each various individual
and collective combinations and permutation of these may not be
explicitly disclosed, each is specifically contemplated and
described herein, for all methods and systems. This applies to all
aspects of this application including, but not limited to, steps in
disclosed methods. Thus, if there are a variety of additional steps
that can be performed it is understood that each of these
additional steps can be performed with any specific embodiment or
combination of embodiments of the disclosed methods.
[0029] Unless stated to the contrary, a formula with chemical bonds
shown only as solid lines and not as wedges or dashed lines
contemplates each possible isomer, e.g., each enantiomer,
diastereomer, and meso compound, and a mixture of isomers, such as
a racemic or scalemic mixture.
[0030] Renal cell carcinoma (RCC) is the most common type of kidney
cancer. It accounts for more than 90% of malignant kidney tumours.
Renal cell carcinoma consists of a heterogenous group of tumours
with distinct genetic and metabolic defects and histopathologic and
clinical features.
[0031] Bezafibrate is a fibric acid hypolipidemic agent chemically
related to clofibrate. Because of its effectiveness in lowering
VLDL cholesterol levels, it is used in the treatment of
hyperlipidemias associated with high triglyceride levels. Like
other fibrates, bezafibrate binds to the peroxisome proliferating
activating receptor alpha (PPAR.alpha.) triggering alterations in
cholesterol levels. Bezafibrate is well absorbed following oral
administration. Bezafibrate is chemically represented:
##STR00001##
[0032] Bezafibrate is currently available under the trade name
Bezalipx 200 mg tablets and Bezalip.RTM. SR 400 mg tablets. The
recommended dosage is one Bezalip.RTM. 200 mg tablet thrice a day
equivalent to 600 mg of bezafibrate or Bezalip.RTM. SR 400 mg
sustained release tablet once daily equivalent to 400 mg of
bezafibrate.
[0033] Bezafibrate is known to stimulate the expression of genes
involved in fatty acid and lipoprotein metabolism thus resulting in
a shift from hepatic fat synthesis to fat oxidation. It reduces
triglycerides by 30-50% and moderately reduce (by 15-20%)
low-density lipoprotein cholesterol (LDL-C). It also leads to a
substantial reduction in serum triglycerides and an increase in
high-density lipoprotein cholesterol levels. It is a synthetic pan
peroxisome proliferator-activated receptor (PPAR) ligand agonist
with increased specificity for PPAR.alpha. as compared to
PPAR.beta., PPAR.delta., and PPAR.gamma..
[0034] Further, PPAR.gamma. is a nuclear receptor that regulates
lipid homeostasis and is implicated in the pathology of numerous
diseases, including cancer such as colon cancer, breast cancer,
lung cancer and prostate cancer as well as inflammations.
[0035] The inventors of the present invention have found that
bezafibrate optionally in combination with other anti-cancer agents
exhibits a significant role in the inhibition of tumor growth,
progression and metastasis of renal cell carcinoma.
[0036] As used herein, term "bezafibrate" is denoted in broad sense
to include not only bezafibrate per se but also its
pharmaceutically acceptable derivatives. Suitable pharmaceutically
acceptable derivatives include pharmaceutically acceptable salts,
pharmaceutically acceptable solvates, pharmaceutically acceptable
hydrates, pharmaceutically acceptable anhydrates, pharmaceutically
acceptable enantiomers, pharmaceutically acceptable esters,
pharmaceutically acceptable polymorphs, pharmaceutically acceptable
esters, pharmaceutically acceptable tautomers, pharmaceutically
acceptable complexes etc.
[0037] Bezafibrate may be formulated as a pharmaceutically
acceptable salt. Pharmaceutically acceptable salts are salts that
retain the desired biological activity of the parent compound and
do not impart undesirable toxicological effects. Examples of such
salts are acid addition salts formed with inorganic acids, for
example, hydrochloric, hydrobromic, sulfuric, phosphoric, and
nitric acids and the like; salts formed with organic acids such as
acetic, oxalic, tartaric, succinic, maleic, fumaric, gluconic,
citric, malic, methanesulfonic, ptoluenesulfonic,
napthalenesulfonic, and polygalacturonic acids, and the like; salts
formed from elemental anions such as chloride, bromide, and iodide;
salts formed from metal hydroxides, for example, sodium hydroxide,
potassium hydroxide, calcium hydroxide, lithium hydroxide, and
magnesium hydroxide; salts formed from metal carbonates, for
example, sodium carbonate, potassium carbonate, calcium carbonate,
and magnesium carbonate; salts formed from metal bicarbonates, for
example, sodium bicarbonate and potassium bicarbonate; salts formed
from metal sulfates, for example, sodium sulfate and potassium
sulfate; and salts formed from metal nitrates, for example, sodium
nitrate and potassium nitrate. Pharmaceutically acceptable and
non-pharmaceutically acceptable salts may be prepared using
procedures well known in the art, for example, by reacting a
sufficiently basic compound such as an amine with a suitable acid
comprising a physiologically acceptable anion. Alkali metal (for
example, sodium, potassium, or lithium) or alkaline earth metal
(for example, calcium) salts of carboxylic acids can also be
made.
[0038] Bezafibrate may be formulated as pharmaceutically acceptable
prodrug, for instance as an ester. Prodrugs can substantially
increase the bioavailability of the compounds, permitting more
effective oral therapy. In some embodiments, the prodrug is a
C.sub.1-C.sub.10 alkyl ester of the 1-carboxylic acid, which may or
may not be substituted. A preferred substituent is carbonyl-oxy and
alkyloxy-carbonyloxy. Exemplary esters include methyl, ethyl,
2-morpholinylethyl, pivaloyloxy-methyl ester,
1-(isopropyloxy-carbonyloxy)ethyl ester, and 1-(acetyloxy)ethyl
ester.
[0039] Bezafibrate can be administered according to various dosing
regimens. For instance, bezafibrate can be administered once a day,
twice a day, three times per day, or even more than three times a
day. Bezafibrate can be administered such that the total daily dose
is at least 50 mg, at least 100 mg, at least 250 mg, at least 500
mg, at least 750 mg, at least 1,000 mg, at least 1,250 mg, at least
1,500 mg, at least 1,750 mg, or at least 2,000 mg. In some
instances, the total daily dose can be from 5-5,000 mg, 10-5,000
mg, 25-5,000 mg, 50-5,000 mg, 100-5,000 mg, 200-2,500 mg, 500-2,500
mg, 10-2,500 mg, 50-2,500 mg, 100-2,500 mg, 100-2,000 mg, 100-1,750
mg, 100-1,500 mg, 100-1,250 mg, 100-1,000 mg, 250-1,500 mg,
250-2,000 mg or 500-2,000 mg. In other embodiments, bezafibrate can
be administered less than once daily, instance, once every two
days, once every three days, once every five days, once every seven
days, once every ten days, once every fourteen days, once every
twenty-eight days or once every month.
[0040] In some instances, bezafibrate can be administered
intermittently, for instance for a period of 1-10 days, followed by
a period in which no bezafibrate is administered (e.g., 1-10 days),
followed by another period e.g., 1-10 days, in which bezafibrate is
administered. The on/off dosing schedule can be repeated as many
times as necessary.
[0041] According to the present invention there is provided a
pharmaceutical composition comprising bezafibrate with one or more
pharmaceutically acceptable excipients for the treatment of renal
cell carcinoma (RCC). Preferably, bezafibrate may be provided in
the form of a pharmaceutical composition such as, but not limited
to, solid unit dosage forms including tablets, capsules (filled
with powders, pellets, beads, mini-tablets, pills, micro-pellets,
small tablet units, multiple unit pellet systems (MUPS),
disintegrating tablets, dispersible tablets, granules, microspheres
and multiparticulates), sachets (filled with powders, pellets,
beads, mini-tablets, pills, micro-pellets, small tablet units,
MUPS, disintegrating tablets, dispersible tablets, granules,
microspheres and multiparticulates), powders for reconstitution and
sprinkles, however, other dosage forms such as controlled release
formulations, lyophilized formulations, modified release
formulations, delayed release formulations, extended release
formulations, pulsatile release formulations, dual release
formulations and the like may fall within the scope of the
invention. Apart from this, it will be well acknowledged by person
skilled in the art to have other forms of pharmaceutical
compositions like liquid or semisolid dosage form (liquids,
suspensions, solutions, dispersions, ointments, creams, emulsions,
microemulsions, sprays, spot-on), injection/parenteral
preparations, topical, inhalations, buccal, nasal etc. and which
may be envisaged under the ambit of the invention.
[0042] Depending on the pathological stage, patient's age and other
physiological parameters, size of the tumor, and the extent of
invasion, the pharmaceutical composition comprising bezafibrate may
require specific dosage amounts and specific frequency of
administrations. Preferably, on an average, the dose range that may
be feasible for producing suitable anticancer effect may range from
25 mg to 3 gms depending on the above factors, and the route of
administration adopted for administering the pharmaceutical
composition. The dosing frequency that may be required for
adherence to the therapy may be at least once, twice or thrice a
day depending on the above mentioned factors and the route of
administration adopted for administering the pharmaceutical
composition.
[0043] It will further be well acknowledged by person skilled in
the art that by specific treatment with bezafibrate, various
physicochemical properties could be improved such as solubility,
better absorption, bioavailability, increased shelf life, etc. and
wherein such specific treatment refers to one or more of
micronization and nanosizing techniques which may achieve one or
more of the benefits aimed hereinabove, and may also assist in dose
reduction. For instance, bezafibrate may be present in the form of
nanoparticles which have an average particle size of less than
2,000 nm, less than 1,500 nm, less than 1,000 nm, less than 750 nm,
less than 500 nm, or less than 250 nm.
[0044] Suitable pharmaceutically acceptable excipients may be used
for formulating the dosage forms according to the present invention
such as, but not limited to, surface stabilizers or surfactants,
viscosity modifying agents, polymers including extended release
polymers, stabilizers, disintegrants or super disintegrants,
diluents, plasticizers, binders, glidants, lubricants, sweeteners,
flavoring agents, anti-caking agents, opacifiers, anti-microbial
agents, antifoaming agents, emulsifiers, buffering agents, coloring
agents, carriers, fillers, anti-adherents, solvents, taste-masking
agents, preservatives, antioxidants, texture enhancers, surface
stabilizers, channeling agents, coating agents or combinations
thereof.
[0045] The present inventors have discovered the bezafibrate is
surprisingly effective for the treatment of renal cell carcinoma.
In certain embodiments, bezafibrate can be used to treat
conventional (clear-cell) renal cell carcinoma, papillary renal
cell-carcinoma, chromophobe renal carcinoma, onco-cytoma, or
collecting-duct carcinoma. Renal cell carcinoma can be classified
in stages, according to the extent of disease progression. The TNM
(tumor size/lymph node/metastasis) system includes the following
stages of RCC:
[0046] Stage I: Tumor of a diameter of 7 cm (approx. 23/4 inches)
or smaller, and limited to the kidney, with no lymph node
involvement or metastases to distant organs.
[0047] Stage II: Tumor larger than 7.0 cm but still limited to the
kidney, with no lymph node involvement or metastases to distant
organs.
[0048] Stage III: Tumor of any size with involvement of a nearby
lymph node but no metastases to distant organs. Tumor of this stage
may be with or without spread to fatty tissue around the kidney,
with or without spread into the large veins leading from the kidney
to the heart; or Tumor with spread to fatty tissue around the
kidney and/or spread into the large veins leading from the kidney
to the heart, but without spread to any lymph nodes or other
organs; or Tumor with spread to fatty tissue around the kidney
and/or spread into the large veins leading from the kidney to the
heart, but without spread to any lymph nodes or other organs.
[0049] Stage IV: Tumor that has spread directly through the fatty
tissue and the fascia ligament-like tissue that surrounds the
kidney; or involvement of more than one lymph node (near or distant
from kidney); or distant metastases, such as in the lungs, bone, or
brain.
[0050] Bezafibrate can be used to treat Stage I RCC, Stage II RCC,
Stage III RCC, or Stage IV RCC. In some embodiments, bezafibrate
can be administered in an amount effective to reduce tumor size,
inhibit tumor growth, alleviate symptoms, delay progression,
prolong survival, including, but not limited to disease free
survival, prevent or delay RCC metastasis, reduce or eliminate
preexisting RCC metastasis, and/or prevent recurrence of RCC. All
of these effects fall within the general scope of treating RCC.
[0051] As used herein, the term "delay" refers to methods that
reduce the probability of disease development/extent in a given
time frame, when compared to otherwise similar methods that do not
include the use of bezafibrate. Probabilities can be established
using clinical trials, but can also be determined using in vitro
assays when correlations have been established. In some
embodiments, bezafibrate can inhibit renal cancer cell
proliferation. For instance, at least about 10%, at least about
20%, at least about 30%, at least about 40%, at least about 50%, at
least about 60%, at least about 70%, at least about 80%, at least
about 90%, or 100% of cell proliferation is inhibited upon
administration of a therapeutically effective amount of
bezafibrate. In some embodiments, bezafibrate can inhibit renal
cancer metastasis. For instance, at least about 10%, at least about
20%, at least about 30%, at least about 40%, at least about 50%, at
least about 60%, at least about 70%, at least about 80%, at least
about 90%, or 100% of metastasis is inhibited upon administration
of a therapeutically effective amount of bezafibrate. Bezafibrate
can also be used to reduce tumor volume. For instance, tumor volume
can be reduced by at least about 10%, at least about 20%, at least
about 30%, at least about 40%, at least about 50%, at least about
60%, at least about 70%, at least about 80%, or at least about 90%
of tumor volume can be reduced upon administration of a
therapeutically effective amount of bezafibrate.
[0052] According to the present invention, there is provided a
method of alleviating or treating renal cell carcinoma (RCC) by
administration of bezafibrate in combination with one or more
anti-cancer drugs either simultaneously, sequentially, or
separately. In certain embodiments, bezafibrate can be administered
with:
[0053] (A) cytotoxic anti-neoplastic drugs such as nucleoside
analogues, antifolates, antimetabolites, topoisomerase I inhibitor,
anthracyclines, podophyllotoxins, taxanes, vinca alkaloids,
alkylating agents, platinum compounds, proteasome inhibitors,
nitrogen mustards & oestrogen analogue; and/or
[0054] (B) targeted anti-neoplastic drugs such as monoclonal
antibodies, tyrosine kinase inhibitors, mTOR inhibitors, retinoids,
immunomodulatory agents, histone deacetylase inhibitors, other
kinase inhibitors.
[0055] In some embodiments, bezafibrate may be administered
(simultaneously, sequentially or separately) with one or more
anti-cancer drugs. Such drugs include small molecule chemical
agents and biological agents, including immunotherapies. Exemplary
anti-cancer drugs include Abiraterone acetate, Methotrexate,
Paclitaxel Albumin-stabilized Nanoparticle, Brentuximab Vedotin,
Ado-Trastuzumab Emtansine, Doxorubicin Hydrochloride, Afatinib
Dimaleate, Everolimus, Netupitant, Palonosetron Hydrochloride,
Imiquimod, Aldesleukin, Alectinib, Alemtuzumab, Melphalan
Hydrochloride, Melphalan, Pemetrexed Disodium, Chlorambucil,
Aminolevulinic acid, Anastrozole, Aprepitant, Pamidronate Disodium,
Exemestane, Nelarabine, Arsenic Trioxide, Ofatumumab, Asparaginase
Erwinia chrysanthemi, Atezolizumab, Bevacizumab, Axitinib,
Azacitidine, Carmustine, Belinostat, Bendamustine hydrochloride,
Bevacizumab, Bexarotene, Tositumomab, Bicalutamide, Bleomycin,
Blinatumomab, Blinatumomab, Bortezomib, Bosutinib, Busulfan,
Cabazitaxel, Cabozantinib, Alemtuzumab, Irinotecan hydrochloride,
Capecitabine, Fluorouracil, Carboplatin, Carfilzomib, Bicalutamide,
Lomustine, Ceritinib, Daunorubicin Hydrochloride, Cetuximab,
Chlorambucil, Cyclophosphamide, Clofarabine, Cobimetinib,
Dactinomycin, Cobimetinib, Crizotinib, Ifosfamide, Ramucirumab,
Cytarabine, Dabrafenib, Dacarbazine, Decitabine, Daratumumab,
Dasatinib, Daunorubicin hydrochloride, Decitabine, Efibrotide
Sodium, Defibrotide sodium, Degarelix, Denileukin Diftitox,
Denosumab, Dexamethasone, Dexrazoxane hydrochloride, Dinutuximab,
Docetaxel, Doxorubicin Hydrochloride, Dacarbazine, Rasburicase,
Epirubicin hydrochloride, Elotuzumab, Oxaliplatin, Eltrombopag
olamine, Aprepitant, Elotuzumab, Enzalutamide, Epirubicin
Hydrochloride, Cetuximab, Eribulin Mesylate, Vismodegib, Erlotinib
hydrochloride, Etoposide, Raloxifene hydrochloride, Melphalan
hydrochloride, Toremifene, Panobinostat, Fulvestrant, Letrozole,
Filgrastim, Fludarabine phosphate, Flutamide, Methotrexate,
Pralatrexate, Recombinant HPV Quadrivalent Vaccine, Recombinant HPV
Nonavalent vaccine, Obinutuzumab, Gefitinib, Gemcitabine
hydrochloride, Gemtuzumab Ozogamicin, Afatinib Dimaleate, Imatinib
Mesylate, Glucarpidase, Goserelin acetate, Eribulin mesylate,
Trastuzumab, Topotecan hydrochloride, Palbociclib, Ibritumomab
tiuxetan, Ibrutinib, Ponatinib hydrochloride, Idarubicin
hydrochloride, Idelalisib, Imiquimod, Axitinib, Recombinant
Interferon Alfa-2b, Tositumomab, Ipilimumab, Gefitinib, Romidepsin,
Ixabepilone, Ixazomib Citrate, Ruxolitinib phosphate, Cabazitaxel,
Ado-Trastuzumab Emtansine, Palifermin, Pembrolizumab, Lanreotide
Acetate, Lapatinib ditosylate, Lenalidomide Lenvatinib mesylate,
Leuprolide acetate, Olaparib, Vincristine Sulfate, Procarbazine
hydrochloride, Mechlorethamine hydrochloride, Megestrol Acetate,
Trametinib, Mercaptopurine, Temozolomide, Mitoxantrone
hydrochloride, Plerixafor, Busulfan, Azacitidine, Gemtuzumab
Ozogamicin, Vinorelbine tartrate, Necitumumab, Nelarabine,
Sorafenib tosylate, Nilotinib, Ixazomib citrate, Nivolumab,
Romiplostim, Obinutuzumab, Ofatumumab, Olaparib, Omacetaxine
mepesuccinate, Pegaspargase, Ondansetron hydrochloride,
Osimertinib, Panitumumab, Panobinostat, Peginterferon Alfa-2b,
Pembrolizumab, Pertuzumab, Plerixafor, Pomalidomide, Ponatinib
hydrochloride, Necitumumab, Pralatrexate, Procarbazine
hydrochloride, Aldesleukin, Denosumab, Ramucirumab, Rasburicase,
Regorafenib, Lenalidomide, Rituximab, Rolapitant hydrochloride,
Romidepsin, Ruxolitinib phosphate, Siltuximab, Dasatinib, Sunitinib
malate, Thalidomide, Dabrafenib, Osimertinib, Talimogene,
Atezolizumab, Temsirolimus, Thalidomide, Dexrazoxane hydrochloride,
Trabectedin, Trametinib, Trastuzumab, Lapatinib ditosylate,
Dinutuximab, Vandetanib, Rolapitant hydrochloride, Bortezomib,
Venetoclax, Crizotinib, Enzalutamide, Ipilimumab, Trabectedin,
Ziv-Aflibercept, Idelalisib, and Ceritinib. Preferred agents for
use in combination with bezafibrate include bevacizumab, axitinib,
sorafenib, sunitinib, everolimus, temsirolimus, pazopanib, and
lenvatinib. An especially preferred agent is sunitinib
[0056] Bezafibrate can be administered with one or more
chemotherapeutic agents either simultaneously, sequentially, or
separately. In certain cases, bezafibrate can be administered for a
period of at least 1 week, at least 2 weeks, at least 4 week, at
least 6 weeks, at least 8 week, or at least 10 weeks, prior to
commencing treatment with additional agents. In some instances,
bezafibrate and the other agent can be administered intermittently,
for instance a period of bezafibrate administration, followed by a
period in which the other agent to administered, followed by
another period of bezafibrate administration. The cycle can be
repeated as many times as necessary.
[0057] In certain cases, the combination of bezafibrate and
additional agent will exhibit a greater than additive effect (i.e.,
a synergistic effect). In other instance, the use of bezafibrate
permits a reduced amount of the other agent to be administered,
without a corresponding decrease in therapeutic efficiency.
[0058] In cases of combination therapy, it is possible that a
unitary dosage form comprising both bezafibrate and one or more
additional anti-cancer drugs may be employed. In some instances,
the combinations may be provided in form of kit including a unitary
dosage form containing both bezafibrate and at least one other
therapeutic agent, or a kit having bezafibrate and an additional
agent in separate dosage forms. In some embodiments, bezafibrate is
present in an oral or parenteral composition and the additional
anti-cancer drug therapy may be provided in an oral or parenteral
composition. In one embodiment, the kit preparation may be provided
in an all oral dosage form presentation wherein both the
bezafibrate and the additional anti-cancer drug are presented in an
oral dosage form. In another embodiment, the kit preparation may be
provided as an oral plus parenteral dosage form presentation
wherein bezafibrate is presented in an oral form and the additional
anti-cancer drug is presented in a parenteral form. Alternatively,
the kit preparation may be provided wherein bezafibrate is
presented in a parenteral form and the additional anti-cancer drug
is presented in an oral dosage form.
[0059] In some instances, bezafibrate can be used in combination
with ionizing radiation and/or surgical interventions for the
treatment of RCC. Bezafibrate can be administered before, during,
or after treatment with ionizing radiation or surgical
intervention. In certain cases, bezafibrate can be administered for
a period of at least 1 week, at least 2 weeks, at least 4 week, at
least 6 weeks, at least 8 week, or at least 10 weeks, prior to
commencing treatment with ionizing radiation or surgery. Exemplary
forms of radiation include x-rays, gamma rays, electron beams and
proton beams. It has been found that administration of bezafibrate
permits a reduction in the total exposure of the patient to
ionizing radiation, without a corresponding reduction in
therapeutic efficiency. In certain instances, bezafibrate can be
administered both prior and subsequent to ionizing radiation and/or
surgical interventions. For instance, bezafibrate can be
administered for a period of at least 1 week, at least 2 weeks, at
least 4 week, at least 6 weeks, at least 8 week, or at least 10
weeks, following treatment with ionizing radiation or surgery.
EXAMPLES
[0060] The following examples are set forth below to illustrate the
methods and results according to the disclosed subject matter.
These examples are not intended to be inclusive of all aspects of
the subject matter disclosed herein, but rather to illustrate
representative methods, compositions, and results. These examples
are not intended to exclude equivalents and variations of the
present invention, which are apparent to one skilled in the
art.
[0061] Efforts have been made to ensure accuracy with respect to
numbers (e.g., amounts, temperature, etc.) but some errors and
deviations should be accounted for. Unless indicated otherwise,
parts are parts by weight, temperature is in .degree. C. or is at
ambient temperature, and pressure is at or near atmospheric. There
are numerous variations and combinations of reaction conditions,
e.g., component concentrations, temperatures, pressures, and other
reaction ranges and conditions that can be used to optimize the
product purity and yield obtained from the described process. Only
reasonable and routine experimentation will be required to optimize
such process conditions.
Example 1: In Vitro 2D Assay
[0062] Cell lines RXF 1183L, RXF 1220L, RXF 1781L, RXF 393L and RXF
486L were established at Oncotest from the corresponding human
patient-derived xenograft. The cell lines 786-O, A-498, Caki-1 and
SN12C were purchased from NCI (National Cancer Institute, Bethesda,
Md., USA). ACHN was purchased from ECACC (European Collection of
Cell Cultures, Salisbury, UK). Authenticity of cell lines was
confirmed at the DSMZ by STR (short tandem repeat) analysis, a PCR
based DNA-fingerprinting methodology.
[0063] Cell lines were routinely passaged once or twice weekly and
maintained in culture for up to 20 passages. All cells were grown
at 37.degree. C. in a humidified atmosphere with 5% CO.sub.2 in
RPMI 1640 medium (25 mM HEPES, with L-glutamine, #FG1385, Biochrom,
Berlin, Germany) supplemented with 10% (v/v) fetal calf serum
(Sigma, Taufkirchen, Germany) and 0.1 mg/mL gentamicin (Life
Technologies, Karlsruhe, Germany).
[0064] Bezafibrate was dissolved at a concentration of 100 mM in
100% Methanol and next 1:5 diluted with cell culture media (20 mM,
20% Methanol). Starting with this solution, serial dilutions
(1:1.6) were prepared with cell culture medium. Next, 150 .mu.L of
the serial dilutions were transferred to the assay plates upon
removing the cell culture media of each well (complete change of
medium on assay plates). Thus the final test concentrations were
10,240, 6,400, 4,000, 2,500, 1,563, 977, 610, and 381 .mu.M. The
maximum methanol concentrations were 6% and 10%. No precipitation
was observed.
[0065] Cells were harvested from exponential phase cultures,
counted and plated in 96-well flat-bottom microtiter plates at a
cell density of 4,000-12,000 cells/well depending on the cell
line's growth rate. After a 24 hour recovery period to allow the
cells to resume exponential growth, test compounds were added.
Compounds were applied at 8 to 10 concentrations in duplicate and
treatment continued for 96 hour. After 96 hour treatment of cells,
20 .mu.L/well CellTiter-Blue.RTM. reagent was added. Following an
incubation period of up to four hours, fluorescence (FU) was
measured by using the Enspire Multimode Plate Reader (excitation
.lamda.=531 nm, emission .lamda.=615 nm).
[0066] For calculations, the mean values of duplicate/quadruplicate
(untreated control) data were used. Sigmoidal
concentration-response curves were fitted to the data points (T/C
values) obtained for each cell line using 4 parameter non-linear
curve fit.
[0067] Bezafibrate was tested in two independent runs by using
different ranges of concentrations. In the first run bezafibrate
was tested in the range from 93 to 4,000 .mu.M, in the second run
from 381 to 10,240 .mu.M. In both runs bezafibrate displayed a
concentration-dependent activity with sigmoidal
concentration-effect curves in all cell lines tested with a
geometric mean absolute IC.sub.50 value of 1,776 .mu.M and 1,222
.mu.M, respectively. In the first run individual IC50 values were
in the range from 1,069 .mu.M (ACHN) to 3,412 .mu.M (1220L) and in
the second run between 795 .mu.M (786-O) and 2,122 .mu.M (A-498)
(FIG. 1).
[0068] Overall, concentration dependent anti-cancer activity with
IC50 values in the low mM range was detected for bezafibrate.
TABLE-US-00001 TABLE 1 Relative and Absolute IC.sub.50 values for
Bezafibrate across the human tumor cell models for RCC in in vitro
2D model. Cell line Absolute IC.sub.50 .mu.M Relative IC.sub.50
.mu.M 1183 960 1056 1220 1930 2106 1781 1862 1946 393 1037 1642 486
987 1328 786-O 795 1073 A-498 2122 2277 ACHN 1051 1127 Caki-1 1229
1398 SN12C 966 1206 Geometric mean 1222 1559
Example 2: In Vitro 3D Assay
[0069] Cell lines were routinely passaged one or twice weekly. All
cells were grown at 37.degree. C. in a humidified atmosphere with
5% CO.sub.2 in RPMI 1640 medium (Biochrom) supplemented with 10%
(v/v) fetal calf serum and 0.1 mg/mL gentamicin. The percentage of
viable cells was determined in a Neubauer-hemocytometer using
trypan blue exclusion.
[0070] Tumor xenografts (patient-derived, as well as cell
line-derived xenografts) were passaged as subcutaneous xenografts
in NMRI nu/nu mice. At a tumor volume of 400-1,000 mm.sup.3
tumor-bearing mice were sacrificed and tumors were collected under
sterile conditions without delay according to the relevant Oncotest
SOPs and the relevant animal welfare guidelines published by the
FELASA and the GV-SOLAS. Tumors were mechanically disaggregated and
subsequently incubated with an enzyme cocktail consisting of
collagenase type IV (41 U/mL), DNase I (125 U/mL), hyaluronidase
type III (100 U/mL), and dispase II (1.0 U/mL) in RPMI 1640 medium
(Life Technologies) at 37.degree. C. for 60-120 minutes. Cells were
passed through sieves of 100 .mu.m and 40 .mu.m mesh size (Cell
Strainer, BD Falcon.TM.), and washed with RPMI 1640 medium. The
percentage of viable cells was determined in a
Neubauer-hemocytometer using trypan blue exclusion. Aliquots of the
cells were frozen down, and stored in liquid nitrogen. On each day
of an experiment, a frozen aliquot of tumor cells was thawed and
used for preparation of assay plates.
[0071] The clonogenic assay was carried out in a 96 well plate
format using ultra low attachment plates. For each test, cells were
prepared as described above and assay plates were prepared as
follows: each test well contained a layer of semi-solid medium with
tumor cells (50 .mu.L), and a second layer of medium supernatant
with or without test compound (100 .mu.L). The cell layer consisted
of 2.510.sup.3 to 110.sup.4 tumor cells per well, which were seeded
in 50 .mu.L/well cell culture medium (IMDM, supplemented with 20%
(v/v) fetal calf serum, 0.01% (w/v) gentamicin, and 0.4% (w/v)
agar). After 24 hours the test compounds were added after serial
dilution in cell culture medium, and left on the cells for the
duration of the experiment (continuous exposure, 100 .mu.l drug
overlay). Every plate included six untreated control wells and
drug-treated groups in duplicate at 9 concentrations. Cultures were
incubated at 37.degree. C. and 7.5% CO.sub.2 in a humidified
atmosphere for 8 to 13 days and monitored closely for colony growth
using an inverted microscope. Within this period, ex vivo tumor
growth led to the formation of colonies with a diameter of >50
.mu.m. At the time of maximum colony formation, counts were
performed with an automatic image analysis system (CellInsight NXT,
Thermo Scientific). 48 hours prior to evaluation, vital colonies
were stained with a sterile aqueous solution of
2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyltetrazolium chloride (1
mg/ml, 100 .mu.l/well).
[0072] Bezafibrate inhibited colony formation in a
concentration-dependent manner with a mean relative IC.sub.50 value
of 985 .mu.M (mean absolute IC.sub.50 value=945 .mu.M). Bottom
plateaus of the concentration-effect curves of responding tumor
models were in the range from 0 to 18%, indicating clear inhibition
of tumor colony growth in the selected test range. Based on
relative IC.sub.50 values, above average activity was observed for
bezafibrate against SN12C cells (IC.sub.50=463 .mu.M) (FIG. 2).
TABLE-US-00002 TABLE 2 Relative and Absolute IC.sub.50 values for
Bezafibrate across the human tumor cell models for RCC in in vitro
3D model. Cell line Absolute IC.sub.50 .mu.M Relative IC.sub.50
.mu.M 1183 609 712 1220 1162 1232 1781 1029 1104 393 1270 1276 486
993 962 786-O 1336 1474 A-498 764 773 ACHN 1013 1057 Caki-1 1287
1290 SN12C 463 463 Geometric mean 945 985
Example 3: In Vitro 3D Combination Assay
[0073] The objective of the study was to assess anti-tumor efficacy
of bezafibrate in combination with sunitinib in a 5.times.5 matrix
combination format against tumor cell lines of renal cancer using a
clonogenic assay with image analysis as read-out. The Bliss
independence methodology was used for data analysis, in order to
identify possible synergistic effects.
[0074] The clonogenic assay was carried out in a 96 well plate
format using ultra low attachment plates. The assay plates were
prepared as follows: each test well contained a layer of semi-solid
medium with tumor cells (50 .mu.l), and a second layer of medium
supernatant with or without test compounds (100 .mu.l). The cell
layer consisted of 3.times.103 to 1.times.104 tumor cells per well,
which were seeded in 50 .mu.l/well cell culture medium (IMDM,
supplemented with 20% (v/v) fetal calf serum, 0.01% (w/v)
gentamicin, and 0.4% (w/v) agar). The soft-agar layer was
immediately covered with 90 .mu.l of the same culture medium
without agar. After 24 hours the test compounds were added after
serial dilution in IMDM and transfer in cell culture medium, and
left on the cells for the duration of the experiment (continuous
exposure, 100 .mu.L total drug overlay). Every plate included six
untreated control wells and drug-treated groups. Cultures were
incubated at 37.degree. C. and 7.5% CO.sub.2 in a humidified
atmosphere for 8 or 13 days and monitored closely for colony growth
using an inverted microscope. Within this period, ex vivo tumor
growth led to the formation of colonies with a diameter of >50
.mu.m. At the time of maximum colony formation, counts were
performed with an automatic image analysis system (CellInsight NXT,
Thermo Scientific). 48 hours prior to evaluation, vital colonies
were stained with a sterile aqueous solution of
2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyltetrazolium chloride (1
mg/ml, 100 .mu.l/well).
[0075] Bezafibrate was tested alone and in combination with
sunitinib in order to investigate the ability to inhibit
anchorage-independent growth and ex vivo colony formation of tumor
cells in semi-solid medium. Bezafibrate was tested against RXF
1183L and RXF SN12C at concentrations ranging from 125 .mu.M to
2000 .mu.M, Sunitinib was tested at concentrations ranging from
0.3125 .mu.M to 5 .mu.M against all cell lines. Results are
depicted in FIGS. 3 and 4.
Example 4: In Vivo Animal Efficacy Study
[0076] Healthy 10 female athymic nude mice were recruited for the
donor cell inoculation. Animals were subcutaneously injected at
flank region with 10 million A498 cells suspended in 200 .mu.l of
media and matrigel. Animals were monitored for solid tumor growth.
Once the tumor reached .about.500 mm.sup.3, donor animals were
humanely sacrificed and tumors were collected under aseptic
condition. Tumors were fragmented in to .about.30 mg size.
[0077] After one week of acclimatization, female athymic nude mice
were subcutaneously implanted with .about.30 mg tumor fragments
using the device trocar. Animals were observed for tumor growth for
next three weeks. Tumor bearing animals were selected from the
experimental animals and grouped on basis of tumor size, into four
groups containing 7 animals in each group as mentioned below.
TABLE-US-00003 Group Treatment Dose, Route & Regimen No. of
Animals G1 Vehicle, 10 mL/kg, p.o. q.d. .times. 21 7 0.5% CMC G2
Sunitinib 20 mg/kg, p.o. q.d. .times. 21 7 G3 Bezafibrate 150
mg/kg, p.o. q.d. .times. 21 7 G4 Bezafibrate + 150 mg/kg, p.o. q.d.
.times. 21 + 7 Sunitinib 20 mg/kg, p.o. q.d. .times. 21
[0078] The tumor sizes were measured weekly twice from the date of
tumor appearance till the end of the experiment. Tumor size was
measured by digital vernier caliper (MITUTOYO) by measuring length
(L=longest axis) and width (W=shortest axis).
[0079] Tumor Volume (V) was calculated using formula:
V=L.times.W2/2 (Unit: mm.sup.3), where L=length of tumor (mm) and
w=width of tumor (mm). Mean tumor volume and % tumor growth
inhibition (% TGI) were then calculated (SEM=standard error of
mean).
Tumor Volume
TABLE-US-00004 [0080] G1 G2 G3 G4 Days Mean SEM Mean SEM Mean SEM
Mean SEM 1 147.56 20.68 145.40 29.19 146.92 19.97 146.87 23.57 6
331.93 79.52 174.90 37.07 198.50 31.03 146.27 38.93 9 463.77 110.07
261.95 66.94 363.14 70.22 179.96 59.99 13 617.07 146.04 340.62
93.64 463.30 111.28 252.51 90.77 16 961.26 226.49 448.05 121.72
709.65 163.77 295.39 110.81 20 1280.51 295.87 630.25 215.72 875.99
206.53 425.61 160.23
Tumor Growth Inhibition (%)
TABLE-US-00005 [0081] Days G1 G2 G3 G4 1 0 1.47 0.44 0.46 6 0 47.31
40.20 55.93 9 0 43.52 21.70 61.20 13 0 44.80 24.92 59.08 16 0 53.39
26.18 69.27 20 0 50.78 31.59 66.76
[0082] Bezafibrate by itself shows anti-tumor efficacy. It inhibits
tumor formation by up to 40% on day 6. Bezafibrate in combination
with sunitinib shows more than additive effect in inhibiting tumor
growth. None of the treatment groups show any significant change in
body weights.
Example 5: Pharmaceutical Compositions
[0083] Manufacturing formula for a pharmaceutical composition
envisaged under the present invention can be referred herein
below:
[0084] Capsules
TABLE-US-00006 Formula I Formula II Formula III Formula IV
Ingredients (mg) (mg) (mg) (mg) Bezafibrate 100-400 100-400 100-400
100-400 Cremophor 150-750 150-750 150-750 150-750 ELP/RH 40 Capmul
MCM 200-500 200-500 200-500 200-500 Labrasol/transcutol -- 50-200
-- -- HP/ethanol Vitamin E TPGS -- -- 10-50 -- Gelucire 44/14 -- --
-- 50-200
1) Cremophor/Gelucire was heated and melted. 2)
Labrasol/transcutol/ethanol was added to the melt obtained in step
(1). 3) Capmul MCM was added to the mixture obtained in step (2) to
form a solution. 4) Vitamin E TPGS was heated and added dropwise to
the solution obtained in step (3) and bezafibrate was added to this
solution. (In examples which do not contain Vitamin E TPGS, only
bezafibrate is added to the solution of step (3). 5) The solution
obtained in step (4) was filled in soft or hard gelatin
capsule.
[0085] Tablets
TABLE-US-00007 Ingredients Qty/Tab (mg) Bezafibrate 200-400
Microcrystalline cellulose 10-35 Lactose 50-200 Croscarmellose
Sodium 2-10 Povidone 3-10 Polysorbate 80 3-10 Methylene
chloride/water q.s. Hypromellose 30-90 Colloidal Anhydrous silica
1-5 Talc 1-5 Magnesium Stearate 1-5 Opadry ready mix 10-20 Purified
water qs
[0086] 1) Bezafibrate, microcrystalline cellulose, lactose and
croscarmellose sodium were blended together to obtain a dry
mix.
[0087] 2) Polysorbate 80 was dissolved in half quantity of
methylene chloride and water to obtain a solution.
[0088] 3) Povidone was added in remaining quantity of methylene
chloride and water and mixed and added to the Polysorbate 80
solution obtained in step (2) to form the binder solution.
[0089] 4) The dry mix obtained in step (1) was granulated with the
binder solution obtained in step (3) to obtain granules.
[0090] 5) The granules obtained in step (4) were dried, sized and
blended with hypromellose, colloidal anhydrous silica and talc.
[0091] 6) The granules obtained in step (5) were lubricated with
magnesium stearate and compressed into tablets and coated.
[0092] The compositions and methods of the appended claims are not
limited in scope by the specific compositions and methods described
herein, which are intended as illustrations of a few aspects of the
claims and any compositions and methods that are functionally
equivalent are intended to fall within the scope of the claims.
Various modifications of the compositions and methods in addition
to those shown and described herein are intended to fall within the
scope of the appended claims. Further, while only certain
representative compositions and method steps disclosed herein are
specifically described, other combinations of the compositions and
method steps also are intended to fall within the scope of the
appended claims, even if not specifically recited. Thus, a
combination of steps, elements, components, or constituents may be
explicitly mentioned herein or less, however, other combinations of
steps, elements, components, and constituents are included, even
though not explicitly stated. The term "comprising" and variations
thereof as used herein is used synonymously with the term
"including" and variations thereof and are open, non-limiting
terms. Although the terms "comprising" and "including" have been
used herein to describe various embodiments, the terms "consisting
essentially of" and "consisting of" can be used in place of
"comprising" and "including" to provide for more specific
embodiments of the invention and are also disclosed. Other than in
the examples, or where otherwise noted, all numbers expressing
quantities of ingredients, reaction conditions, and so forth used
in the specification and claims are to be understood at the very
least, and not as an attempt to limit the application of the
doctrine of equivalents to the scope of the claims, to be construed
in light of the number of significant digits and ordinary rounding
approaches.
* * * * *